The present invention provides a stroboscopic optical image mapping system comprising a control module, an optical module, and an image acquiring unit. The control unit forms a delayed pulse signal by modulating a first pulse signal having a plurality of pulses with a pulse period, wherein a time interval between two adjacent pulses of the delayed pulse signal has a time difference with respect to the pulse period. The optical module provides an incident light to be projected on an organic object, which has a dye therein and is stimulated by a second pulse signal for generating a sequential action potential. The fluorescence generates from the dye inside the organic object, which corresponds to the intensity of the sequential action potential. The image acquiring unit is actuated to acquire the fluorescent light according to the delayed pulse signal, thereby forming a plurality of fluorescent images.